Lullaby reagent

Manufactured by Ozyme

Lullaby reagent is a laboratory chemical used for a specific purpose in research or analysis. It is a colorless liquid with a neutral pH. The reagent is designed to perform a core function, but its intended use requires further details that are not provided in this factual description.

Automatically generated - may contain errors

Lab products found in correlation

Opti mem, by Thermo Fisher Scientific (2 mentions) Pspax2, by Addgene (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions) Lipofectamine 2000 reagent, by Thermo Fisher Scientific (2 mentions) Control sirna, by Horizon Discovery (1 mentions) Fortessa 2 flow cytometer, by BD (1 mentions) Orca r2 camera, by Hamamatsu Photonics (1 mentions) On targetplus smartpool, by Horizon Discovery (1 mentions) L 15 culture medium, by Merck Group (1 mentions) Mda mb 231, by American Type Culture Collection (1 mentions) Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions) Puromycin, by Santa Cruz Biotechnology (1 mentions) Dogtor, by Ozyme (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Puromycin, by Thermo Fisher Scientific (1 mentions) Plko 1 puro lentiviral vector, by Merck Group (1 mentions) Opti mem medium, by Thermo Fisher Scientific (1 mentions) Genejuice, by Merck Group (1 mentions) Glomax multi detection system, by Promega (1 mentions)

Close spelling variants of this product

Lullaby (9 citations) Lullaby transfection reagent (8 citations) Lullaby siRNA transfection reagent (3 citations)

11 protocols using lullaby reagent

PRL Signaling in T47-D Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

The human mammary epithelial cell line T47-D was kindly provided by Dr V. Goffin (INSERM UMR_S1151-CNRS UMR8253, Institut Necker-Enfants Malades, Paris, France) and tested for contamination. Cells were maintained at 5% CO₂ in a 37°C incubator and grown in a phenol red-free RPMI 1640 medium containing 10% FBS and 5 µg/ml insulin, as previously described (Baker et al., 2016 (link)).
For siRNAs knockdown experiments, cells were transfected the day after their seeding with siRNA controls (siLuciferase) or specific siRAB6A/A′ (50 nM final concentration) in Lullaby reagent (OZ Biosciences). A second transfection was performed 24 h after the first one. Experiments were conducted 72 h after the last transfection.
Cells were washed in PBS 24 h before PRL treatment and medium was replaced by a serum-free medium. Cells were treated with human PRL (Sigma Aldrich, #L4021) at 250 ng/ml for 5, 15 and 45 min at 37°C in a medium supplemented with 5 µg/ml insulin and 1 µM dexamethasone, as previously described (Baker et al., 2016 (link)).
Immunofluorescence staining of PRLR (anti-PRLR, Invitrogen; #35-9200; 1/200) and RAB6 (AC306, produced and purified in B.G.’s lab; 1/1000; Goud et al., 1990 (link)) were performed after fixing the cells with 4% paraformaldehyde and mild permeabilization with 0.05% saponin.

Cayre S., Faraldo M.M., Bardin S., Miserey-Lenkei S., Deugnier M.A, & Goud B. (2020). RAB6 GTPase regulates mammary secretory function by controlling the activation of STAT5. Development (Cambridge, England), 147(19), dev190744.

+ Open protocol

+ Expand

siRNA Knockdown and Plasmid Transfection

Check if the same lab product or an alternative is used in the 5 most similar protocols

Hela cells were transfected for 72 hr using Lullaby reagent (OZ biosciences) with siRNA from Dharmacon against Capns1 (#L-009979–00), UBC9 (#L-004910–00), SAE2 (#L-005248–01), RhoGDIα (#L-016253–00) or control siRNA (#D-001810–10, Dharmacon) according to the manufacturers’instructions. The pEGFP-RhoGDIα WT or pEGFP-RhoGDIα K138R expression vectors were, respectively, a kind gift from Dr. Mark R. Philips (New York University School of Medicine, New York) and Dr. Chuanshu Huang (New York University School of Medicine, New York). HeLa cells were transfected with plasmids by using Lipofectamine 2000 (Invitrogen) according to the manufacturer’s protocol.

Lapaquette P., Fritah S., Lhocine N., Andrieux A., Nigro G., Mounier J., Sansonetti P, & Dejean A. (2017). Shigella entry unveils a calcium/calpain-dependent mechanism for inhibiting sumoylation. eLife, 6, e27444.

+ Open protocol

+ Expand

Silencing ARF6 in MDA-MB-231 cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

siRNA transfection was performed using 50 nM siRNA with Lullaby reagent (OZ Biosciences) according to manufacturer’s instructions and analyzed 72 hrs after treatment. The siRNAs used in this study are listed in S2 Table.
For lentivirus production, HEK293T cells were transfected using GeneJuice (Novagen) with a mix of ARF6-T157N expression vector and psPAX2 (AddGene) and pVSV-G (Clonetech) packaging vectors in OPTIMEM (Invitrogen). After 72 hrs, virus-containing supernatant was collected, filtered and used for transduction of a subconfluent monolayer of MDA-MB-231 cells.

Marchesin V., Montagnac G, & Chavrier P. (2015). ARF6 Promotes the Formation of Rac1 and WAVE-Dependent Ventral F-Actin Rosettes in Breast Cancer Cells in Response to Epidermal Growth Factor. PLoS ONE, 10(3), e0121747.

+ Open protocol

+ Expand

Homologous Recombination and NHEJ Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols

Reporter cell lines were used to assess homologous recombination (DR-GFP) and non-homologous end joining (EJ5-GFP) [13] (link). For transfections, I-SceI expression vector (pCBASce), GFP expression vector (pCAGGS-NZEGFP) and a control empty vector (pCAGGS-BSKX) were used [13] (link). In detail, 2 × 10⁵ cells were plated into a 12-well dish and transfected with non-targeting or siRNA targeting ATMIN (Dharmacon) using Lullaby reagent (OZ Biosciences). After 48 h, cells were transfected with I-SceI or control plasmids. Transfection complexes were prepared by mixing Lipofectamine 2000 (Life Technologies) in OptiMEM (Invitrogen) with 0.8 μg of expression vectors for I-SceI or control vectors per sample. Samples were analyzed 3 days after transfection by immunoblotting to assess the knockdown efficiency. The frequency of GFP⁺ cells was determined on a Fortessa II flow cytometer (BD Bioscience).

Schmidt L., Wiedner M., Velimezi G., Prochazkova J., Owusu M., Bauer S, & Loizou J.I. (2014). ATMIN is required for the ATM-mediated signaling and recruitment of 53BP1 to DNA damage sites upon replication stress. DNA Repair, 24, 122-130.

+ Open protocol

+ Expand

High-Throughput siRNA Screening of Breast Cancer

Cited 4 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

The gene library for the siRNA screen was chosen from a meta-analysis of breast cancer published sequencing studies (1 (link),12 (link)–16 (link)), choosing the top 200 recurrently mutated genes that encompassed the top 100 genes in ER+, HER2+ and TNBC respectively (Table S1). Reverse transfection siRNA spheroid screens were performed as previously described in triplicate (5 ,11 (link)). MCF10a, MCF10NeoT, MCF10AT1, MCF10DCIS.com, MCF10Ca1a, MCF10Ca1d, MCF10Ca1h cell lines were reverse-transfected with 37.5nM of Dharmacon siGENOME siRNA using Lullaby reagent (Oz biosciences). BT20, CAL-51, HCC70, HCC1937, HCC1806, Hs578t and MDA-MB-157 cell lines were reverse-transfected with 37.5nM of Dharmacon siGENOME siRNA using Viromer (lipocalyx). Cell viability was measured after 5 days by CellTiter-Glo. The progression series screen was analysed using z-score analysis. The TNBC cell line targeted screen was analysed by plate median normalised values. Spheroids were imaged using a Nikon TE 2000 inverted wide field microscope fitted with a motorised stage, filter wheels and a Pro-Scan controller (Prior Scientific), Shutter (Sutter instruments), Orca R2 camera (Hamamatsu), 84000v2 DAPI/FITC/TRITC/Cy5 Quad (Chroma technology). The microscope is operated by HCI imaging software 4.3.1.33. Experiments are carried out at 37⁰C (Solent Scientific) and 10% CO₂.

Peck B., Bland P., Mavrommati I., Muirhead G., Cottom H., Wai P.T., Maguire S.L., Barker H.E., Morrison E., Kriplani D., Yu L., Gibson A., Falgari G., Brennan K., Farnie G., Buus R., Marlow R., Novo D., Knight E., Guppy N., Kolarevic D., Susnjar S., Milijic N.M., Naidoo K., Gazinska P., Roxanis I., Pancholi S., Martin L.A., Holgersen E.M., Cheang M.C., Noor F., Postel-Vinay S., Quinn G., McDade S., Krasny L., Huang P., Daley F., Wallberg F., Choudhary J.S., Haider S., Tutt A.N, & Natrajan R. (2021). 3D functional genomics screens identify CREBBP as a targetable driver in aggressive triple-negative breast cancer. Cancer research, 81(4), 847-859.

+ Open protocol

+ Expand

MDA-MB-231 Cell Line Characterization

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The human breast adenocarcinoma cell line MDA-MB-231 (American Type Culture Collection, ATCC HTB-26) was maintained in L-15 culture medium (Sigma-Aldrich) with 2 mM glutamine and 15% FCS at 37 °C in 1% CO₂. MDA-MB-231 cells stably expressing MT1-MMPmCh were cultured in the presence of 0.5 mg/mL G41843 (link). MDA-MB-231 cells were transfected with plasmid constructs by using Amaxa nucleotransfection. Cells were analyzed 24 h after transfection. Small inhibitory RNAs targeting MT1-MMP (MMP14, L-004145-00) were SMARTpoolON-TARGETplus from Dharmacon. Cortactin (CTTN, SI02661960 and SI02662485), LIMK1 (LIMK1, SI00605542 and SI00605549) and LIMK2 (LIMK2, SI02665334 and SI02758490) siRNAs were purchased from Qiagen. Cells were treated with specific siRNA (50 nM final concentration) with Lullaby reagent (OZ Biosciences) and analyzed 72 h after treatment.

Lagoutte E., Villeneuve C., Lafanechère L., Wells C.M., Jones G.E., Chavrier P, & Rossé C. (2016). LIMK Regulates Tumor-Cell Invasion and Matrix Degradation Through Tyrosine Phosphorylation of MT1-MMP. Scientific Reports, 6, 24925.

+ Open protocol

+ Expand

Cell line engineering and siRNA depletion

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

DLD1, HEK293T and HeLa cell lines were obtained from the Cell Services Science Technology Platform at the Francis Crick Institute. Cells were cultured in DMEM (Thermo Fisher Scientific) with 10% FCS (Thermo Fisher Scientific). DLD1-εWT, DLD1-εD383/451 N and DLD1-εD536N have been engineered from the original cell lines DLD1-FRT-Trex (kindly provided by Prof. Stephen Taylor). Cells were depleted of endogenous protein by siRNA and cultured in DMEM containing 10% FCS and tetracycline (100 ng/ml) for 24 h prior to assay. Cells were treated with inhibitors for the time/s stated. Cell lines were routinely tested for mycoplasma. Cell synchronisation was performed as previously described^{4 (link)}. Unless otherwise indicated, HEK293T cells were transiently transfected with purified plasmid DNA using the Lipofectamine 2000 reagent (Thermo), for 24–48 h prior to analysis. siRNA transfection was performed using the Lullaby reagent (Oz Biosciences) and the siRNAs listed in Supplementary Table 2 were purchased from Dharmacon and used at 20 nM.

Martini S., Davis K., Faraway R., Elze L., Lockwood N., Jones A., Xie X., McDonald N.Q., Mann D.J., Armstrong A., Ule J, & Parker P.J. (2021). A genetically-encoded crosslinker screen identifies SERBP1 as a PKCε substrate influencing translation and cell division. Nature Communications, 12, 6934.

+ Open protocol

+ Expand

Luciferase Reporters for NMD Regulation In Vivo

Cited 2 times

Check if the same lab product or an alternative is used in the 5 most similar protocols

In order to express luciferase NMD reporters in vivo, HeLa cells (grown to 60–80% confluency) were transfected with the reporter constructs in pcDNA 3.1 (+) vectors (described above). Short hairpin RNA (shRNA)-mediated RNA interference (RNAi) degradation⁵⁵ was applied to knockdown UPF1 as described previously^{35 (link),53 (link),55}. The transfection mix consisted of 20 ng/µl reporter plasmid, 20 ng/µl pSUPuro plasmid (1:1 mixture of pSUPuro UPF1 against two target sequences^{53 (link)}) in Opti-MEM containing 3% (v/v) Dogtor (OZ Biosciences) transfection reagent. As a negative control, a pSUPuro plasmid containing a randomized target sequence was used (pSUPuro Scr). After 12 h the medium was replaced by DMEM +/+ containing 1.5 µg/ml Puromycin (Santa Cruz Biotechnology). The antibiotic selection was carried out for 48 h until the medium was replaced with DMEM +/+ to let the cells recover for approx. 24 h. To obtain a list of NMD-sensitive RNAs 3 × 10⁵ HeLa cells per well were seeded in 6-well plates. Twenty-four hours later, the cells were transfected with 52 pmol of siRNA using Lullaby reagent (OZ Biosciences). After 48 h, the cells were re-transfected as before. Protein and total RNA were isolated after one additional day. The siRNA sequence 5′-GAUGCAGUUCCGCUCCAUU-3′ was used for targeting UPF1.

Karousis E.D., Gurzeler L.A., Annibaldis G., Dreos R, & Mühlemann O. (2020). Human NMD ensues independently of stable ribosome stalling. Nature Communications, 11, 4134.

+ Open protocol

+ Expand

Transfection and Serum Modulation of Cancer Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Breast and prostate cancer cells were reverse-transfected with 37.5 nM of Dharmacon siGENOME siRNA using Lullaby reagent (Oz biosciences). After 24 h, culture medium was replaced with either 10 % (full serum) or 1 % (low serum) foetal calf serum (FCS) containing medium. Additional supplementations were included for the experiments indicated. After 72 h, cells were fixed in 80 % ethanol over night at −20 °C. Plates were subsequently stained with DAPI (Sigma), and cell number was determined using the ACUMEN X3 microplate cytometer.

Peck B., Schug Z.T., Zhang Q., Dankworth B., Jones D.T., Smethurst E., Patel R., Mason S., Jiang M., Saunders R., Howell M., Mitter R., Spencer-Dene B., Stamp G., McGarry L., James D., Shanks E., Aboagye E.O., Critchlow S.E., Leung H.Y., Harris A.L., Wakelam M.J., Gottlieb E, & Schulze A. (2016). Inhibition of fatty acid desaturation is detrimental to cancer cell survival in metabolically compromised environments. Cancer & Metabolism, 4, 6.

+ Open protocol

+ Expand

siRNA and shRNA Knockdown Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols

siRNA treatments were performed using 50 nM siRNA (see Table S5 for the list of siRNAs) and Lullaby reagent (OZ Biosciences) according to the manufacturer’s instructions and analyzed 72 h after treatment. shRNAs against human ARF6 and MT1-MMP inserted in pLKO.1-puro lentiviral vector were purchased from Sigma-Aldrich (Table S5). For lentivirus production, HEK293T cells were transfected using GeneJuice (EMD Millipore) with a mix of expression vector and psPAX2 (Addgene) and pVSV-G (Takara Bio Inc.) packaging vectors in OPTIMEM medium (Invitrogen). After 72 h, the virus-containing supernatant was collected, filtered, and used for transduction of a subconfluent monolayer of MDA-MB-231 or MCF10DCIS.com cells. 1 µg/ml puromycin (Gibco) was added after 48 h for selection.

Marchesin V., Castro-Castro A., Lodillinsky C., Castagnino A., Cyrta J., Bonsang-Kitzis H., Fuhrmann L., Irondelle M., Infante E., Montagnac G., Reyal F., Vincent-Salomon A, & Chavrier P. (2015). ARF6–JIP3/4 regulate endosomal tubules for MT1-MMP exocytosis in cancer invasion. The Journal of Cell Biology, 211(2), 339-358.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!